Procyanidin B2 Alleviates Heat-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells
Abstract
:1. Introduction
2. Results
2.1. PB2 Ameliorates Acute HS-Induced Cell Viability Loss
2.2. PB2 Alleviates Oxidative Stress Induced by HS
2.3. PB2 Protects against HS-Induced Inflammatory Response
2.4. PB2 Further Promotes the Activation of Nrf2/HO-1 Signaling Pathway
2.5. Nrf2 Knockdown and Its Effect on MAC-T Cell Viability
2.6. PB2 Alleviates HS-Induced Oxidative Stress and Inflammatory Response through the Nrf2 Signaling Pathway
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatment
4.2. Cell Viability Assay
4.3. Antioxidant Capacity Measurement
4.4. Detection of ROS
4.5. Determination of IL-1β and TNF-α
4.6. Transient Transfection of Nrf2
4.7. RNA Isolation and Quantitative Real-Time PCR
4.8. Immunofluorescence Staining
4.9. Protein Extraction and Western Blotting
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | GenBank ID | Primer Sequences (5′ to 3′) | Product Size |
---|---|---|---|
HSP70 | NM_203322.3 | Forward: CAAGATCAGCGAGGCGGACAAG Reverse: ACACCTGCTCCAGCTCCTTCC | 128 bp |
TNF-α | NM_173966.3 | Forward: CTGGCGGAGGAGGTGCTCTC Reverse: GGAGGAAGGAGAAGAGGCTGAGG | 85 bp |
IL-1β | NM_174093.1 | Forward: ATGAAGAGCTGCATCCAACACCTG Reverse: ACCGACACCACCTGCCTGAAG | 110 bp |
Nrf2 | NM_001011678.2 | Forward: TCAGCCAGCACAACACATACCATC Reverse: ACGGGAATGTCTCTGCCAAAAGC | 128 bp |
HO-1 | NM_001014912.1 | Forward: CCGCTACCTGGGAGACCTGTC Reverse: ACTTGGTGGCACTGGCGATATTG | 128 bp |
Txnrd1 | NM_174625.5 | Forward: CGTGCCTTACATCTATGCCATCGG Reverse: TGGAGCCACCATACAGCCTCTG | 110 bp |
GAPDH | NM_001034034.2 | Forward: TGCCCGTTCGACAGATAGCC Reverse: GCGACGATGTCCACTTTGCC | 148 bp |
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Wang, H.; Hao, W.; Yang, L.; Li, T.; Zhao, C.; Yan, P.; Wei, S. Procyanidin B2 Alleviates Heat-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells. Int. J. Mol. Sci. 2022, 23, 7769. https://doi.org/10.3390/ijms23147769
Wang H, Hao W, Yang L, Li T, Zhao C, Yan P, Wei S. Procyanidin B2 Alleviates Heat-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells. International Journal of Molecular Sciences. 2022; 23(14):7769. https://doi.org/10.3390/ijms23147769
Chicago/Turabian StyleWang, Hongzhuang, Weiguang Hao, Liang Yang, Tingting Li, Chongchong Zhao, Peishi Yan, and Shengjuan Wei. 2022. "Procyanidin B2 Alleviates Heat-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells" International Journal of Molecular Sciences 23, no. 14: 7769. https://doi.org/10.3390/ijms23147769